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COMPARISON OF IN VIVO AND IN VITRO CHEMICAL SCREENING ASSAYS FOR ESTROGEN-RESPONSIVE PROTEIN BIOMARKER EXPRESSION IN THE SHEEPSHEAD MINNOW (CYPRINODON VARIEGATUS)
Citation:
WILKINSON, S. S., J. NG, P. S. HARRIS, K. SALINAS, C. C. WALKER, AND M. J. HEMMER. COMPARISON OF IN VIVO AND IN VITRO CHEMICAL SCREENING ASSAYS FOR ESTROGEN-RESPONSIVE PROTEIN BIOMARKER EXPRESSION IN THE SHEEPSHEAD MINNOW (CYPRINODON VARIEGATUS). Presented at SETAC North America 28th Annual Meeting, Milwaukee, WI, November 11 - 15, 2007.
Impact/Purpose:
abstract for presentation
Description:
Animal welfare concerns and the high cost associated with in vivo chemical screening methods have prompted efforts to develop highly specific and sensitive in vitro techniques to evaluate chemicals for endocrine activity. Diagnostic biomarkers developed using in vitro methods should be associated with expression of identical biomarkers, specific toxicity pathways, or adverse effects in whole animal studies. In this study, a comparison of the presence and relative abundance of four estrogen-responsive protein biomarkers discovered in sheepshead minnows (SHM) plasma was made with protein expression in a SHM hepatocyte in vitro assay. Plasma from male SHM treated for 7 days to aqueous concentrations of 0.05, 0.1, 0.2, 0.5 and 1.0 upg/L of 17B-estradiol (E2),, and growth media from cultures of female SHM hepatocytes treated with media concentrations of 13.5 — 1,350 upg E2/L for 4 days were examined using matrix assisted laser desorption and ionizations-time of flight (MALDI-TOF) mass spectrometry. Four estrogen-responsive protein biomarkers at 2950, 2978, 3003 and 3025 m/z were significantly elevated in both the in vivo and in vitro E2 treatments although the hepatocyte media assay demonstrated a lower relative abundance of the four biomarkers. A clear dose-associated increase in biomarker expression was evident with increasing E2 concentration in both assay systems.